Recombinant production of a highly efficient photolyase from Thermus thermophilus

• DOI: 10.1002/biot.202300325
• Publication Date: 2024-02-22T12:25:46Z
• Authors: Karin Torres‐Obreque; Felipe Gobbi Gonç...; Rafael Bertelli F...; Fabiana Fuentes‐León; Carlos Frederico ...; Tales Alexandre C...; Gisele Monteiro; Patrizia Perego; Carlota de Olivei...
• Keywords: Thermus thermophilus; Photolyase; Photoprotection

Abstract Ultraviolet (UV) radiation from sunlight can damage DNA, inducing mutagenesis and eventually leading to skin cancer. Topical sunscreens are used avoid the effect of UV irradiation, but topical application DNA repair enzymes, such as photolyase, provide active photoprotection by recovery. Here we produced a recombinant Thermus thermophilus photolyase expressed in Escherichia coli , evaluated kinetic parameters bacterial growth kinetics stability enzyme. The maximum biomass (𝑋 𝑚𝑎𝑥 ) 2.0 g L −1 was reached after 5 h cultivation, corresponding 𝑃 X = 0.4 h. µ corresponded 1.0 . Photolyase purified affinity chromatography high amounts pure enzyme were obtained (3.25 mg cultivation). Two different methods demonstrated activity on samples very low concentrations, 15 µg mL already resulted 90% CPD photodamage removal. We also determined k M 9.5 nM, confirming potential at conservation freezing (‐20°C) lyophilization. Therefore, demonstrate T. capacity its an ingredient be incorporated dermatological products.